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Nuclear Envelope Formation In Vitro: A Sea Urchin Egg Cell-Free System

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The Nucleus

Part of the book series: Methods in Molecular Biology ((MIMB,volume 464))

Abstract

The formation of the nuclear envelope (NE) typically occurs once during every mitotic cycle in somatic cells, and also around the sperm nucleus following fertilization. Much of our understanding of NE assembly has been derived from systems modeling the latter event in vitro. In these systems, demembranated sperm nuclei are combined with fertilized egg cytoplasmic extracts and an ATP-regenerating system and in a multistep process they form the functional double bilayer of the NE. Using a system that we developed from sea urchin gametes, we have demonstrated that NE assembly is regulated by membrane vesicles in a spatial and temporal fashion, emphasizing the roles of phosphoinositides, particularly phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2), diacylglycerols (DAG), and lipid-modifying enzymes in NE assembly.

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Acknowledgments

This work was supported by an Amherst College Faculty Research Award of the H. Axel Schupf ‘57 Fund for Intellectual Life (DLP) and core funding from Cancer Research UK (BL).

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© 2008 Humana Press, a part of Springer Science + Business Media, LLC

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Byrne, R.D., Zhendre, V., Larijani, B., Poccia, D.L. (2008). Nuclear Envelope Formation In Vitro: A Sea Urchin Egg Cell-Free System. In: Hancock, R. (eds) The Nucleus. Methods in Molecular Biology, vol 464. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-461-6_12

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  • DOI: https://doi.org/10.1007/978-1-60327-461-6_12

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60327-460-9

  • Online ISBN: 978-1-60327-461-6

  • eBook Packages: Springer Protocols

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